LOCUS       sequence-414391-        6166 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-AIA expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6166)
  AUTHORS   Williams-Jones DP, Webby MN, Press CE, Gradon JM, Armstrong SR, 
            Szczepaniak J, Kleanthous C
  TITLE     Tunable force transduction through the Escherichia coli cell 
            envelope.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Nov 21;120(47):e2306707120. doi: 
            10.1073/pnas.2306707120. Epub 2023 Nov 16.
  PUBMED    37972066
REFERENCE   2  (bases 1 to 6166)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-11-21"; volume: "120(47):e2306707120. 
            doi"; pages: " 10.1073/pnas.2306707120. Epub 2023 Nov 16"
COMMENT     Sequence Label: pDWJ26
FEATURES             Location/Qualifiers
     source          1..6166
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(96..974)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1001..1285
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1208..1227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1363..2076
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1363..2076
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1393..1421)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2016..2036
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     CDS             2077..2106
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="ASMTGGQQMG"
     primer_bind     complement(3024..3041)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3194..3280
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3372..3399
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3418..3509
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3510..3578,3579..4370)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3728..3747)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4412..4867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4499..4518)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4709..4730
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      4978..5566
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5467..5486
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5720..5737
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5903..5925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6063..6082
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atcgatgcat aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac
       61 tccgtcaagc cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca
      121 ttcacttttt cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta
      181 aatacccgcg agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata
      241 ggcatccggg tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag
      301 cttaagacgc taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag
      361 caaacatgct gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg
      421 tactgacaag cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct
      481 tccatgcgcc gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc
      541 ccttcccctt gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc
      601 gcttcatccg ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca
      661 tgccagtagg cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga
      721 tgacgaccgt agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa
      781 acaaattctc gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata
      841 taacctttca ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc
      901 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      961 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
     1021 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
     1081 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
     1141 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
     1201 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
     1261 tcgcaactct ctactgtttc tccatacccg tttttttggg ctagcaggag gaattcacca
     1321 tggtacccgg ggatcctcta gatttaagaa ggagatatac atatgagtaa aggagaagaa
     1381 cttttcactg gagttgtccc aattcttgtt gaattagatg gtgatgttaa tgggcacaaa
     1441 ttttctgtca gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt
     1501 atttgcacta ctggaaaact acctgttcca tggccaacac ttgtcactac tttcgcgtat
     1561 ggtcttcaat gctttgcgag atacccagat catatgaaac agcatgactt tttcaagagt
     1621 gccatgcccg aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac
     1681 aagacacgtg ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa
     1741 ggtattgatt ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac
     1801 tcacacaatg tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa
     1861 attagacaca acattgaaga tggaagcgtt caactagcag accattatca acaaaatact
     1921 ccaattggcg atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc
     1981 ctttcgaaag atcccaacga aaagagagac cacatggtcc ttcttgagtt tgtaacagct
     2041 gctgggatta cacatggcat ggatgaacta tacaaagcta gcatgactgg tggacagcaa
     2101 atgggtcgcg gatccgtgtc aaaggcaacc gaacaaaacg acaagctcaa gcgggcgata
     2161 attatttcag cagtgctgca tgtcatctta tttgcggcgc tgatctggag ttcgttcgat
     2221 gagaatatag aagcttcagc cggaggcggc gcggattata aaaccggtgg caacggtaat
     2281 ggtggcccga cgggtagcgg taacagcggt ggcggtggca gtggcaatgg tggcggtaat
     2341 ggcaatagtg gtggcggcag cggcaccggc ggcaatggta gcgcgggcaa tggtaacggt
     2401 ggtccgaccg gcagcggtaa tagcggtggt ggcggtagtg gcaacggcgg tggtaacggt
     2461 aacagcggcg gcggtagtgg tacgggcggc aatggcagcg ccggtaacgg caacggtggt
     2521 ccaaccggca gtggtaatag tggtggtggt ggcagcggta atggtggtgg caacggcaac
     2581 agtggcggtg gtagcggcac gggcggtaac ggcagcgcgc ccgggaccgg tgggggaggg
     2641 gcgaaaggga acaatgcttc gcctgccggg agtggtaata ctaaaaacaa tggcgcatca
     2701 ggggccgata tcaataacta tgccgggcag attaaatctg ctatcgaaag taagttctat
     2761 gacgcatcgt cctatgcagg caaaacctgt acgctgcgca taaaactggc acccgatggt
     2821 atgttactgg atatcaaacc tgaaggtggc gatcccgcac tttgtcaggc tgcgttggca
     2881 gcagctaaac ttgcgaagat cccgaaacca ccaagccagg cagtatatga agtgttcaaa
     2941 aacgcgccat tggacttcaa accgtaagtc gacctgcagg catgcaagct tggctgtttt
     3001 ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag aagcggtctg
     3061 ataaaacaga atttgcctgg cggcagtagt gcggtggtcc cacctgaccc catgccgaac
     3121 tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc gagagtaggg
     3181 aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     3241 ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag cggatttgaa
     3301 cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa ctgccaggca
     3361 tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac aaactctttt
     3421 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3481 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     3541 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     3601 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     3661 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     3721 aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag caactcggtc
     3781 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     3841 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     3901 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     3961 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     4021 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     4081 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     4141 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     4201 ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg gggccagatg
     4261 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     4321 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     4381 aagtttactc atatatactt tagattgatt tacgcgccct gtagcggcgc attaagcgcg
     4441 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
     4501 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     4561 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     4621 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     4681 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     4741 aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     4801 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     4861 acaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     4921 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     4981 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5041 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5101 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     5161 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5221 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5281 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5341 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5401 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5461 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5521 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5581 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     5641 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     5701 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     5761 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atagggtcat ggctgcgccc
     5821 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5881 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5941 ccgaaacgcg cgaggcagaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc
     6001 accataccca cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca
     6061 tcggtgatgt cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc
     6121 acgatgcgtc cggcgtagag gatctgctca tgtttgacag cttatc
//
